Alterations of lectin binding during chondrogenesis of mouse limb buds

Abstract

The binding of six different FITC-labelled lectins to mesenchyme, blastemal cells and cartilage was investigated in limb buds of mouse embryos during their development from day 10 to day 13. Concanavalin A, wheat germ agglutinin and phaseolus vulgaris agglutinin labelled mesenchymal cells of earlier stages, day 10 or 11, distinctly more than those of later stages. Chondrogenic blastema, basement membrane and muscle were always strongly stained. The galactosamine-specific ricinus communis agglutinin (RCA) bound preferentially to the blastema, whereas the mesenchyme was only weakly labelled. The galactose-specific peanut agglutinin (PNA), however, stained solely the blastema. In the mesenchyme, no binding was detectable light microscopically with this lectin. In cartilage, RCA- and PNA-staining was found to a lesser extent. With the fucose-specific lectin Lotus A, no staining was detectable. Due to the apparent differences in the binding of PNA in mesenchyme and blastema, peroxidase-labelled PNA was used to study the binding behaviour electron microscopically. It is shown that peroxidase-PNA very strongly labelled the intercellular matrix and the plasma membrane of cells in the late blastemal stages, whereas in young blastema no reaction product was detectable. In contrast to light microscopic findings, some label could be demonstrated also in the mesenchyme. The results show a general reduction of lectin receptors in more developed mesenchyme of later stages and the occurrence of galactose and galactose derivatives during early chondrogenesis in the matrix and at the cell membrane. The significance of these changes for chondrogenesis, however, remains to be elucidated.